title: The tethering complex HOPS catalyzes assembly of the soluble SNARE Vam7 into fusogenic trans-SNARE complexes.
abstract: The fusion of yeast vacuolar membranes depends on the disassembly of cis-soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes and the subsequent reassembly of new SNARE complexes in trans. The disassembly of cis-SNARE complexes by Sec17/Sec18p releases the soluble SNARE Vam7p from vacuolar membranes. Consequently, Vam7p needs to be recruited to the membrane at future sites of fusion to allow the formation of trans-SNARE complexes. The multisubunit tethering homotypic fusion and vacuole protein sorting (HOPS) complex, which is essential for the fusion of vacuolar membranes, was previously shown to have direct affinity for Vam7p. The functional significance of this interaction, however, has been unclear. Using a fully reconstituted in vitro fusion reaction, we now show that HOPS facilitates membrane fusion by recruiting Vam7p for fusion. In the presence of HOPS, unlike with other tethering agents, very low levels of added Vam7p suffice to induce vigorous fusion. This is a specific recruitment of Vam7p rather than an indirect stimulation of SNARE complex formation through tethering, as HOPS does not facilitate fusion with a low amount of a soluble form of another vacuolar SNARE, Vti1p. Our findings establish yet another function among the multiple tasks that HOPS performs to catalyze the fusion of yeast vacuoles.
authors.affiliation: Department of Biochemistry, Geisel School of Medicine at Dartmouth, Hanover, NH 03755-3844.
authors.forename: Michael
authors.initials: M
authors.lastname: Zick
authors.forename: William
authors.initials: W
authors.lastname: Wickner
mesh_terms: D055162:Biocatalysis; D019921:Biotinylation; D007295:Inositol Phosphates / Q000378:metabolism; D046912:Multiprotein Complexes / Q000378:metabolism*; D050600:SNARE Proteins / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D012995:Solubility; D019809:Streptavidin / Q000378:metabolism; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism*
chemical_list: D007295:Inositol Phosphates; D046912:Multiprotein Complexes; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; D050825:Synaptosomal-Associated Protein 25; C494205:VAM7 protein, S cerevisiae; C052128:inositol 3-phosphate; D019809:Streptavidin